Reverse transcriptases and uses thereof

Abstract

Hybrid reverse transcriptases formed from portions of FLVRT and MLVRT are provided.

What is claimed is: 
     
       1. A nucleic acid comprising a polynucleotide encoding a hybrid reverse transcriptase, the hybrid reverse transcriptase comprising a finger domain, a palm domain, a thumb domain, a connection domain and an RNase H domain, wherein
 (A) the hybrid reverse transcriptase comprises: 
 (i) a portion of feline leukemia virus reverse transcriptase (FLVRT) comprising the finger domain and the palm domain, wherein the portion of the FLVRT is at least 95% identical to SEQ ID NO:26, said portion of the FLVRT linked to a portion of mouse leukemia virus reverse transcriptase (MLVRT) comprising the thumb domain, the connection domain, and the RNase H domain, wherein the portion of the MLVRT is at least 95% identical to SEQ ID NO:28; or 
 (ii) a portion of feline leukemia virus reverse transcriptase (FLVRT) comprising the finger domain, the palm domain, the thumb domain, and the connection domain wherein the portion of the FLVRT is at least 95% identical to SEQ ID NO:27, said portion of the FLVRT linked to a portion of mouse leukemia virus reverse transcriptase (MLVRT) comprising the RNase H domain, wherein the portion of the MLVRT is at least 95% identical to SEQ ID NO:29; or 
 (B) the hybrid reverse transcriptase comprises: 
 a portion of mouse leukemia virus reverse transcriptase (MLVRT) comprising the finger domain and the palm domain, wherein the portion of the MLVRT is at least 95% identical to SEQ ID NO:1, said portion of the MLVRT linked to 
 a portion of feline leukemia virus reverse transcriptase (FLVRT) comprising the thumb domain, the connection domain, and the RNase H domain, wherein the portion of the FLVRT is at least 95% identical to SEQ ID NO:5. 
 
     
     
       2. The nucleic acid of  claim 1 , comprising
 a portion of feline leukemia virus reverse transcriptase (FLVRT) comprising the finger domain and the palm domain, wherein the portion of the FLVRT is at least 95% identical to SEQ ID NO:26, said portion of the FLVRT linked to 
 a portion of mouse leukemia virus reverse transcriptase (MLVRT) comprising the thumb domain, the connection domain, and the RNase H domain, wherein the portion of the MLVRT is at least 95% identical to SEQ ID NO:28. 
 
     
     
       3. The nucleic acid of  claim 2 , wherein the hybrid reverse transcriptase comprises SEQ ID NO:30. 
     
     
       4. The nucleic acid of  claim 1 ,
 a portion of feline leukemia virus reverse transcriptase (FLVRT) comprising the finger domain, the palm domain, the thumb domain, and the connection domain wherein the portion of the FLVRT is at least 95% identical to SEQ ID NO:27, said portion of the FLVRT linked to a portion of mouse leukemia virus reverse transcriptase (MLVRT) comprising the RNase H domain, wherein the portion of the MLVRT is at least 95% identical to SEQ ID NO:29. 
 
     
     
       5. The nucleic acid of  claim 4 , wherein the hybrid reverse transcriptase comprises SEQ ID NO:31. 
     
     
       6. The nucleic acid of  claim 1 , having at least one mutation that improves thermostability. 
     
     
       7. The nucleic acid of  claim 1 , wherein the hybrid reverse transcriptase comprises:
 a portion of mouse leukemia virus reverse transcriptase (MLVRT) comprising the finger domain and the palm domain, wherein the portion of the MLVRT is at least 95% identical to SEQ ID NO:1, said portion of the MLVRT linked to a portion of feline leukemia virus reverse transcriptase (FLVRT) comprising the thumb domain, the connection domain, and the RNase H domain, wherein the portion of the FLVRT is at least 95% identical to SEQ ID NO:5. 
 
     
     
       8. The nucleic acid of  claim 7 , wherein the portion of the MLVRT comprises SEQ ID NO:1 and the portion of the FLVRT comprises SEQ ID NO:5. 
     
     
       9. The nucleic acid of  claim 8 , wherein the portion of the FLVRT comprises SEQ ID NO:6 or SEQ ID NO:10. 
     
     
       10. The nucleic acid of  claim 9 , wherein the portion of the FLVRT comprises SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:11. 
     
     
       11. The nucleic acid of  claim 8 , wherein the portion of the MLVRT comprises SEQ ID NO:2. 
     
     
       12. The nucleic acid of  claim 8 , wherein the portion of the MLVRT comprises SEQ ID NO:3 or SEQ ID NO:4. 
     
     
       13. The nucleic acid of  claim 8 , wherein:
 the portion of the MLVRT comprises SEQ ID NO:4 and the portion of the FLVRT comprises SEQ ID NO:7; or 
 the portion of the MLVRT comprises SEQ ID NO:4 and the portion of the FLVRT comprises SEQ ID NO:8; or 
 the portion of the MLVRT comprises SEQ ID NO:4 and the portion of the FLVRT comprises SEQ ID NO:9; or 
 the portion of the MLVRT comprises SEQ ID NO:3 and the portion of the FLVRT comprises SEQ ID NO:11; or 
 the portion of the MLVRT comprises SEQ ID NO:3 and the portion of the FLVRT comprises SEQ ID NO:10; or 
 the portion of the MLVRT comprises SEQ ID NO:4 and the portion of the FLVRT comprises SEQ ID NO:13. 
 
     
     
       14. The nucleic acid of  claim 8 , wherein the hybrid reverse transcriptase comprises a sequence at least 95% identical to SEQ ID NO: 14, 15, 16, 17, 18, 19, 34, or 35. 
     
     
       15. The nucleic acid of  claim 1 , comprising a promoter operably linked to the polynucleotide. 
     
     
       16. An expression vector comprising the nucleic acid of  claim 1 . 
     
     
       17. A cell comprising the nucleic acid of  claim 1 .